LamininB4 Cytocaf®
Pack size: 1 mL, 1 mg/mL
Price: £100
Available from TCS Biosciences Ltd.
Features checklist
Recombinant protein polymer that mimics the action of Laminin Beta 4 for adhesive cell culture:
- Provides adhesive surface: Ideal for cells such as iPSCs
- Easy to handle: No need to keep cool, can be used at room temperature and above without denaturing or losing activity
- Simple 1 hour protocol for coating plates
- Free from animal material
Product description
Cytocaf® proteins are based on the bacterial protein Caf1, which associates into long, flexible and bioinert polymers. Bioactivity can be imparted by genetically encoding bioactive motifs into the protein’s sequence. Cytocaf polymers are highly stable, modular and flexible.
LamininB4 Cytocaf® mimics the action of the extracellular matrix protein Laminin (Beta 4 Subunit) through a genetically encoded bioactive peptide that can bind to integrin receptors. This makes it adhesive for a range of cell types including both human and animal iPSCs.
Cells grown on LamininB4 Cytocaf® display adherence and morphologies similar to those seen on recombinant Laminin and Geltrex™ [1].
Integrin/Laminin B4 Interaction
Integrin/LamininB4 Cytocaf® Interaction
References
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Engineering bacterial protein polymers to support human pluripotent stem cell growth and differentiation in culture.
Data & specifications
| Test | Result |
|---|---|
| Bacterial growth | Negative |
| Fungal growth | Negative |
| Particulate examination | Clear |
| Mycoplasma | Negative |
| Host Cell DNA | Undetectable |
Figure 1: (Left) Human iPSCs were seeded onto LamininB4 Cytocaf® coated non tissue culture treated 3.5cm plates and grown for three days in E8 flex media, before imaging with a light microscope at 10X magnification. (Right) Human iPSCs were seeded at a 1 in 8 split ratio into 3.5 cm non-tissue culture treated dished pre-coated with LamininB4 Cytocaf® and grown in E8 flex media supplemented with 10 µM rock inhibitor. After 24 hours, the media was changed and the cells were left to become 80% confluent with media changes performed every other day. Cells were subcultured in the same way. At passage 1, 3 and 5 cells were harvested for RNA analysis. RNA was harvested, reverse transcribed to cDNA and analysed using quantitative real-time PCR. OCT4, SOX2, and NANOG are well-known markers of iPSCs, and CD44 was used as the negative marker.
Figure 2: After 5 passages of human iPSCs on LamininB4 Cytocaf® coated plates, cells were seeded at a split ratio of 1 in 5 into LowAdhere Cytocaf® (which is non-adhesive for iPSCs) coated 6 cm non-TC treated dish in E8 flex media supplemented with 4 mg/mL polyvinyl alcohol (PVA) and 10 µM ROCKi. After 24 hours the media was changed to E6 media with 4 mg/mL PVA. The same media exchange was performed every 2-3 days for 7 days before embryoid bodies were harvested for qPCR analysis.